Das Biswadip, Butler J Scott, Sherman Fred
Department of Biochemistry and Biophysics, University of Rochester School of Medicine and Dentistry, Rochester, New York 14642, USA.
Mol Cell Biol. 2003 Aug;23(16):5502-15. doi: 10.1128/MCB.23.16.5502-5515.2003.
A nuclear mRNA degradation (DRN) system was identified from analysis of mRNA turnover rates in nup116-Delta strains of Saccharomyces cerevisiae lacking the ability to export all RNAs, including poly(A) mRNAs, at the restrictive temperature. Northern blotting, in situ hybridization, and blocking transcription with thiolutin in nup116-delta strains revealed a rapid degradation of mRNAs in the nucleus that was suppressed by the rrp6-delta, rai1-delta, and cbc1-delta deletions, but not by the upf1-delta deletion, suggesting that DRN requires Rrp6p, a 3'-to-5' nuclear exonuclease, the Rat1p, a 5'-to-3' nuclear exonuclease, and Cbc1p, a component of CBC, the nuclear cap binding complex, which may direct the mRNAs to the site of degradation. We propose that certain normal mRNAs retained in the nucleus are degraded by the DRN system, similar to degradation of transcripts with 3' end formation defects in certain mutants.
通过对酿酒酵母nup116-Δ菌株在限制温度下mRNA周转速率的分析,鉴定出一种核mRNA降解(DRN)系统。该菌株在限制温度下缺乏输出所有RNA(包括多聚腺苷酸化mRNA)的能力。在nup116-Δ菌株中进行的Northern印迹、原位杂交以及用硫链丝菌素阻断转录实验表明,细胞核中的mRNA会快速降解,rrp6-Δ、rai1-Δ和cbc1-Δ缺失可抑制这种降解,但upf1-Δ缺失则不能,这表明DRN需要Rrp6p(一种3'至5'核外切核酸酶)、Rat1p(一种5'至3'核外切核酸酶)和Cbc1p(CBC的一个组分,即核帽结合复合体),它们可能将mRNA引导至降解位点。我们提出,细胞核中保留的某些正常mRNA会被DRN系统降解,这类似于某些突变体中具有3'末端形成缺陷的转录本的降解。